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Ultra-Small Gold Nanoparticles by Galvanic Replacement of Larger ULTRA-SMALL GOLD NANOPARTICLES BY GALVANIC REPLACEMENT OF LARGER SILVER NANOPARTICLES by CHANG-TING LIN Presented to the Faculty of the Graduate School of The University of Texas at Arlington in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE THE UNIVERSITY OF TEXAS AT ARLINGTON May 2019 Copyright © by Chang-Ting Lin 2019 All Rights Reserved ACKNOWLEDGMENT I want to thank everyone who has helped me during the past two years, but first I want to send my gratitude to my advisor Dr. Yaowu Hao for his guidance and patience for me to learn many things about nanotechnology and finish my thesis. I would also like to thank my colleagues especially Shahab Ranjbar Bahadori who always provided courage and supports. I am also grateful to the great team in the Radiology Department of UT Southwestern Medical Center in Dallas, Aditi Mulgaokar who significantly contributed to this study. I especially appreciate Dr. Xiankai Sun, whose knowledge and support help this work possible. Finally, I would like to thank my family in Taiwan. They always give me endless support and encouragement for me to finish my master’s degree. ABSTRACT ULTRA-SMALL GOLD NANOPARTICLES BY GALVANIC REPLACEMENT OF LARGER SILVER NANOPARTICLES Chang-Ting Lin, MS The University of Texas at Arlington, 2019 Supervising Professor: Yaowu Hao In the last two decades, Au nanoparticles have been extensively explored for their biomedical applications, which is mainly driven by their biocompatibility and special optical properties known as surface plasmon resonance (SPR) effect. A variety of Au nanoparticles with different sizes and shapes have been synthesized and tested both in vitro and in vivo. For in vivo applications, it is usually required that Au nanoparticles can be cleared through urine (renal clearance). Renal clearance is determined by several factors of Au nanoparticles, including size, shape and surface modification. In this thesis, for the purpose of using Au nanoparticles as carriers to carry radioactive isotopes such as Cu-64 or Pd-103 into human body for cancer imaging and treatment, we developed a process to synthesize, functionalize and be labelled with Cu of ultra small Au nanoparticles. We developed an easy and fast way to synthesize ultra small Au nanoparticles with an average diameter less than 5 nm using galvanic replacement reaction to replace spherical Ag nanoparticles with Au chloroaurate. This synthesis process can be completed within an hour at room temperature. To ensure the renal clearance property, Au nanoparticles with size less than 2 nm have been successfully isolated from synthesized Au nanoparticles. The renal clearance of the nanoparticles are also strongly related to the surface chemistry of the nanoparticles. The Ag nanoparticles we used for the synthesis of Au nanoparticles have polyvinylpyrrolidone (PVP) coating on the surface. PVP is not an ideal surface coating for renal clearance. ii Therefore, we first introduced a simple way to remove the PVP coating by washing the particles with acetone. Then the surface of the Au nanoparticles was functionalized with glutathione which has been proven to be a good surface coating for renal clearance. In addition, we have incorporated Cu on to the surface of these Au nanoparticles using the electroless deposition process, creating a protocol for the radiolabeling of Au nanoparticles. iii TABLE OF CONTENTS ABSTRACT ii TABLE OF CONTENTS iv LIST OF FIGURES vii LIST OF TABLES xi 1. INTRODUCTION 1 2. BACKGROUND INFORMATION 3 2.1 Au Nanoparticles 3 2.1.1 History of Au Nanoparticles 3 2.1.2 Synthesis Methods of Au Nanoparticles with Different Sizes or Shapes 4 2.1.2.1 Au Nanosphere 5 2.1.2.2 Au Nanorod 6 2.1.2.3 Au Nanocage 8 2.1.2.4 Au Nanoshell 10 2.1.2.5 Au Nanoplate 10 2.1.3 Ultra Small Au Nanoparticles 12 2.1.3.1 Optical Property of Au Nanocluster 12 2.1.3.2 Synthesis Method of Au Nanoclusters 12 2.2 Renal Clearable Nanoparticles 13 2.2.1 Introduction to Renal Clearance 13 2.2.2 Properties affecting the renal clearance 14 2.2.3 Renal Clearance of Different Nanoparticles 16 iv 2.2.3.1 Iron Oxide Nanoparticle 16 2.2.3.2 Silicon Nanoparticle 17 2.2.3.3 Au Nanoparticle 18 2.3 Galvanic Replacement Reaction 21 2.4 Electroless Deposition 23 3. SYNTHESIS OF ULTRA-SMALL AU NANOPARTICLES BY GALVANIC REPLACEMENT REACTION OF LARGER AG NANOPARTICLES 25 3.1 Polyvinylpyrrolidone (PVP)-coated Au-Ag Nanoparticles 25 3.1.1 Synthesis of PVP coated Au-Ag NPs by Galvanic Replacement of Big Ag Nanoparticles 25 3.1.2 Characterization of PVP-Coated Au-Ag Nanoparticles 26 3.2 Naked AuNPs 32 3.2.1 Removing PVP Coating of Au Nanoparticles 32 3.2.2 Characterization of Naked Au Nanoparticles 32 3.3 AuNPs Smaller Than 2 nm 33 3.3.1 Isolation of AuNPs smaller than 2 nm 33 3.3.2 Characterization of naked AuNPs smaller than 2 nm 34 3.4 Glutathione (GSH) -coated AuNPs 38 3.4.1 Synthesis 38 3.4.2 Characterization of GSH-coated AuNPs 39 4. INCORPORATION OF CU ONTO AU NANOPARTICLES BY ELECTROLESS DEPOSITION 42 4.1 Mechanism of Cu Electroless Deposition on AuNPs 42 4.2 Electroless Deposition of Cu onto the Surface of the Au Nanoparticles (Cu-Au NPs) 43 v 4.3 Characterization of Cu-Au Nanoparticles (Cu-AuNPs) 44 4.4 Incorporation Rate of Cu onto the AuNPs 46 5. CONCLUSION 49 LIST OF REFERENCE 50 vi LIST OF FIGURES Figure 2. 1 The Lycurgus cup in a) the reflected light and b) transmitted light [19] 4 Figure 2. 2 TEM images of Au nanorods with different aspect ratio: 7.6 (left) and 2.6 (right). 7 Figure 2. 3 Representative electron micrographs of Au nanoparticles and nanorods corresponding to the spectra a±h in Figure 1. Considering only the spheroids and rods, the average aspect ratios of the particles are 1.5, 2.4, 6.1, 8.0, and 10.0 for sets c, d, f, g, and h, respectively. The particles in sets a and b are nearly spherical and have aspect ratios very close to 1.0. 8 Figure 2. 4 (A) SEM of Ag nanocubes. Inset: electron diffraction indicates they are single-crystals. (B) SEM of the product after 0.30 mL of 1 mM HAuCl4 solution was added to a 5-mL 0.8 mM Ag nanocube suspension; a pinhole (lower inset) is observed on the exposed face of ∼1 in 6 nanocubes. Upper inset: TEM of a microtomed sample reveals early hollowing out. (C) SEM of the product after 0.50 mL of HAuCl4 solution was added. Inset: TEM of a microtomed sample reveals the hollow interior of the nanobox. (D) SEM of the product after 2.25 mL of HAuCl4 solution was added. Porous nanocages produced. (E) Illustration summarizing morphological changes. Coloration indicates the conversion of an Ag nanocube into an Au/Ag nanobox then a predominately Au nanocage. [26] 9 Figure 2. 5 (a)–(f) TEM images of nanoshell growth on 120 nm diameter silica dielectric nanoparticle. (a) Initial Au colloid-decorated silica nanoparticle. (b)–(e) Gradual growth and coalescence of Au colloid on the silica nanoparticle surface. (f) Completed growth of metallic nanoshell. [27] 10 Figure 2. 6 Morphological characterization of Au nanoplates on multilayer graphene is grown with KAuBr4 as the Au precursor. The synthesis time t is 5 h. (a, b) Representative scanning electron microscopy (SEM) images of Au nanoplates on multilayer graphene. (c) Typical atomic force microscopy (AFM) image of an Au nanoplate supported on an oxidized Si(001) substrate. The nanoplate edge contrast is enhanced for clarity. (d) Surface height profile measured along the red dashed line in panel c [28]. 11 vii Figure 2. 7 Schematic of the physiology mechanism of the kidney. 14 Figure 2. 8 (a) Ideal disease targeting of renal clearable nanoparticles (NPs) in clinical practices: the NPs specifically target the diseases and untargeted ones are rapidly cleared out of the body through the urinary system. (b) The schematic structure of the kidney corpuscle. (c) Glomerular filtration is a nanoscale phenomenon. The glomerular capillary wall is made of three specialized layers: fenestrated endothelium, glomerular basement membrane, and podocyte extensions of glomerular epithelial cells [36]. 15 Figure 2. 9 The distribution of IONPs in different organs were quantified by ICP-MS at 24 h post-injection [39]. 17 Figure 2.10 a) Schematic diagram depicting the structure and in vivo degradation process for the (biopolymer-coated) nanoparticles used in this study. b) In vivo biodistribution and biodegradation of LPSiNPs over a period of 4 weeks in a mouse. Aliquots of LPSiNPs were intravenously injected into the mouse (n=3 or 4, dose=20 mg kg−1). The silicon concentration in the organs was determined at different time points after injection using ICP-OES [41]. 18 Figure 2.11 Urinalysis of glutathione-coated Au nanoparticles (n = 5 for each time point, 200 μL injections) shows a significant percentage of particles is cleared through renal filtration within 1 h, continues for up to 24 h and returns to baseline concentrations within 1 week. This indicates the glutathione-coated Au nanoparticles is capable of quick passage into the kidneys and bladder and does not cause a strain on the renal system as the injection concentration is increased. (ICP- MS detection limit = 0.4 ppb). Baseline time points (taken before injection) were essentially 0 ppb at all concentrations [46]. 19 Figure 2.12 Organ distribution analysis at 24 h, 2 weeks, and 4 weeks (n = 5 for each time point, 200 μL injections) show an initial accumulation within the kidney and lungs at 24 h.
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